[논문]열수추출 과정에서 삽주, 백출(큰꽃삽주), 북창출 배합이 감초 성분의 추출률에 미치는 영향

김정훈

doi:10.6116/kjh.2019.34.5.39.

열수추출 과정에서 삽주, 백출(큰꽃삽주), 북창출 배합이 감초 성분의 추출률에 미치는 영향
Chemical influences of the rhizomes of Atractylodes japonica, A. macrocephala, or A. chinensis on the extraction efficiencies of chemical compounds in the roots and rhizomes of Glycyrrhiza uralensis during hot-water extraction 원문보기

大韓本草學會誌 = The Korea journal of herbology, v.34 no.5, 2019년, pp.39 - 47

김정훈 (부산대학교 한의학전문대학원 약물의학부)

Abstract ▼ AI-Helper

Objectives : When herbal medicines are extracted together, they may interact with each other, leading to change of chemical characteristics. This study aimed to evaluate the influence of Atractylodes rhizomes (Atractylodes japonica, A. macrocephala, and A. chinensis) on the chemical features of the roots and rhizomes of Glycyrrhiza uralensis, which is are commonly combined with herbal medicines in many herbal formulae, when they are co-decocted. Methods : Liquiritin apioside, liquiritin, ononin, and glycyrrhizin levels of G. uralensis in hot-water extracts prepared by the combination of Atractylodes rhizomes with various weight ratios (G. uralensis : Atractylodes rhizomes = 10:0, 10:5, 10:10, and 10:20) and extraction times (60, 90, and 120 min) were quantified using a HPLC-diode array detector and compared by statistical analysis. Results : The concentrations of liquiritin apioside, liquiritin, ononin, and glycyrrhizin from G. uralensis roots and rhizomes mostly reduced when co-extracted with Atractylodes rhizomes, and the addition of A. chinensis most reduced their contents between Atractylodes combination groups. A. japonica and A. macrocephala rhizomes also showed differences of liquiritin and glycyrrhizin levels at 10 g and 20 g groups of Atractylodes rhizomes. Extraction times also affected the concentrations of liquiritin, ononin, and glycyrrhizin mostly during 60 and 90 min. Conclusions : Atractylodes rhizomes might alter the chemical characteristics of G. uralensis when these herbs are co-decocted. This study provides the understanding of the chemical interactions of herbal medicines during the extraction in hot water.

주제어

표/그림 (9)

그림 Fig. 1. Chemical structures of the marker compounds. 1, Liquiritin apioside; 2, liquiritin; 3, ononin; 4, glycyrrhizin.
그림 Fig. 2. HPLC chromatograms of the marker compounds (A–D) and hot-water extracts of G. uralensis (E), G. uralensis + A. japonica (F),G. uralensis + A. macrocephala (G), and G. uralensis + A. chinensis (H). 1, Liquiritin apioside; 2, liquiritin; 3, ononin; 4, glycyrrhizin.
표 Table 1. Linear equations, correlation coefficients (r²), LODs, and LOQs of the four marker compounds
표 Table 2. Intra- and inter-day precisions of the four marker compounds
그림 Fig. 3. Change in the concentrations of the four marker compounds after hot water co-extraction of G. uralensis (10 g) with each of the three of Atractylodes rhizomes (0, 5, 10, and 20 g) for 120 min. A, Liquiritin apioside; B, liquiritin; C, ononin; D, glycyrrhizin.
표 Table 3: Recoveries of the four marker compounds
그림 Fig. 4. Change in the concentrations of the four marker compounds after hot water co-extraction of G. uralensis (10 g) with Atractylodes rhizomes (10 g) for 60, 90, and 120 min. A, Liquiritin apioside; B, liquiritin; C, ononin; D, glycyrrhizin.
그림 Fig. 5. Regression equations and correlation coefficients (R²)of the four marker compounds in decoction of G. uralensis roots and rhizomes plus Atractylodes rhizomes.
그림 Fig. 6. Regression equations and correlation coefficients (R²)of the four marker compounds in G. uralensis roots and rhizomes (10 g) plus Atractylodes rhizomes (10 g) co-extracts in using different decoction times (60, 90, and 120 min).

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제안 방법

An Agilent 1260 liquid chromatography system (Agilent Technologies, Palo Alto, CA, USA) equipped with an autosampler, degasser, quaternary solvent pump, and diode array detector was used for the quantitative analysis. Data were processed using Chemstation software (Agilent Technologies Inc.
Column, mobile phase modifier, and UV detector wavelength were optimized for HPLC analysis of the four marker compounds in the hot-water extracts of G. uralensis roots and rhizomes. Separation of four marker compounds in G.
Elution was conducted using a gradient elution program as follows: 15% (B) for 0–3 min, 15–40% (B) for 3–35 min, 40% (B) for 1 min, 40–80% (B) for 36–42 min, 80% (B) for 1 min, and then re-equilibrated to 15% (B) until the end of the analysis.
Separation of four marker compounds in G. uralensis extract was performed on a C18–coated silica column (Capcell Pak Mg II; 4.6 ㎜ × 250 ㎜, 5 ㎛; Shiseido).
The accuracy of the HPLC method was determined by testing the recoveries of three known amounts of the six marker compounds (low, medium, and high concentrations) added to sample solutions. Recovery was calculated as follows: Recovery (%) = [(detected concentration – initial concentration) / spiked concentration] × 100.
The precisions were determined by analyzing low, medium, and high concentrations of the abovementioned working solutions three times within a day (intraday precision) and over three consecutive days (interday precision). Precisions are represented as relative standard deviations (RSDs), where RSD (%) = [(standard deviation / mean)×100].

대상 데이터

Elution was conducted using a gradient elution program as follows: 15% (B) for 0–3 min, 15–40% (B) for 3–35 min, 40% (B) for 1 min, 40–80% (B) for 36–42 min, 80% (B) for 1 min, and then re-equilibrated to 15% (B) until the end of the analysis. The diode-array detector was set at ultraviolet wavelengths of 250 and 275 ㎚.
(HPLC grade; Phillipsburg, NJ, USA). Trifluoroacetic acid and liquiritin were purchased from Sigma-Aldrich (St Louis, MO, USA). Liquiritin apioside was purchased from Chengdu Biopurify Phytochemicals Ltd.

데이터처리

Working solutions were injected into the HPLC to construct calibration curves. The linearities of calibration curves were evaluated using correlation coefficients (r²). LOD and LOQ were defined as signal-to-noise (S/N) ratio of 3 and 10, respectively.

성능/효과

1. Co-extraction of G. uralensis with Atractylodes rhizomes reduced the concentrations of liquiritin apioside, liquiritin, ononin, and glycyrrhizin in G. uralensis hot-water extracts and this effect was greatest for co-extraction with A. chinensis.
2. The concentrations of liquiritin apioside, liquiritin, ononin, and glycyrrhizin from G. uralensis roots and rhizomes were altered by the presence of either of three Atractylodes species during hot-water extraction and depended on different weight of Atractylodes rhizomes present and extraction times.
As also shown in figure 3, significant decreases in the concentrations of the four compounds were observed when G. uralensis roots and rhizomes were extracted with A. chinensis (25.878 ± 5.063 ㎍/㎖ at 5 g and 28.671 ± 4.389 ㎍/㎖ at 10 g for liquiritin apioside;25.039 ± 8.777 ㎍/㎖ at 10 g for liquiritin; 1.467 ±0.200 ㎍/㎖ at 5 g and 1.921 ± 0.458 ㎍/㎖ at 10 g for ononin; 117.794 ± 11.992 ㎍/㎖ at 10 g for glycyrrhizin) as compared with extractions in the presence of A. japonica (31.856 ± 8.193 ㎍/㎖ at 5 g and 34.783 ± 6.637 ㎍/㎖ at 10 g for liquiritin apioside; 38.669 ± 7.621 ㎍/㎖ at 10 g for liquiritin;2.126 ± 0.682㎍/㎖ at 5 g and 2.507 ± 0.474 ㎍/㎖ at 10 g for ononin; 162.858 ± 29.785 ㎍/㎖ at 10 g for glycyrrhizin) or A. macrocephala (34.542 ± 7.637㎍/㎖ at 10 g for liquiritin; 1.976 ± 0.457 ㎍/㎖ at 5 g and 2.412 ± 0.387 ㎍/㎖ at 10 g for ononin;163.903 ± 44.960 ㎍/㎖ at 20 g for glycyrrhizin).
Concentrations of liquiritin (26.154 ± 5.360 ㎍/㎖ ↔ 37.533 ± 12.725 ㎍/㎖) and glycyrrhizin (109.861± 12.024 ㎍/㎖ ↔ 163.903 ± 44.960 ㎍/㎖) differed significantly at 20 g groups and those of glycyrrhizin (162.858 ± 29.785 ㎍/㎖ ↔ 134.437 ± 30.683 ㎍/㎖) different at 10 g groups, comparing the presences of A. japonica and A. macrocephala rhizomes.
Concentrations of ononin and glycyrrhizin were significantly reduced by co-extraction with A. japonica for 90 min (2.425 ± 0.294 ㎍/㎖ → 1.630 ± 0.516 ㎍/㎖ for ononin; 149.535 ± 19.512 ㎍/㎖ → 118.239 ±37.466 ㎍/㎖ for glycyrrhizin) and A. chinensis (2.425± 0.294 ㎍/㎖ → 1.517 ± 0.262 ㎍/㎖ for ononin at 90 min; 107.993 ± 24.315 ㎍/㎖ → 91.283 ± 5.901㎍/㎖ for glycyrrhizin at 60 min).
Significant reductions in the concentrations of liquiritin apioside (37.037 ±7.049 ㎍/㎖ → 30.151 ± 4.190 ㎍/㎖, 29.905 ± 6.864㎍/㎖, and 26.522 ± 3.496 ㎍/㎖) and ononin (2.547 ±0.558 ㎍/㎖ → 1.976 ± 0.457 ㎍/㎖ and 2.000 ± 0.301㎍/㎖) occurred in the presence of A. macrocephala rhizomes at all weights except 10 g.
The recovery of the marker compounds in the HPLC methods ranged from ere recovered from 99.89% to 114.61% with RSDs of < 7.7% (Table 3).

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표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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